Objective:

2

To explore the potential effects and mechanisms of Liang-Ge-San (LGS) for the treatment of acute respiratory distress syndrome (ARDS) through network pharmacology analysis and to verify LGS activity through biological experiments.

Methods:

2

The key ingredients of LGS and related targets were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. ARDS-related targets were selected from GeneCards and DisGeNET databases. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed using the Metascape Database. Molecular docking analysis was used to confirm the binding affinity of the core compounds with key therapeutic targets. Finally

the effects of LGS on key signaling pathways and biological processes were determined by

in vitro

and

in vivo

experiments.

Results:

2

A total of LGS-related targets and 496 ARDS-related targets were obtained from the databases. Network pharmacological analysis suggested that LGS could treat ARDS based on the following information: LGS ingredients luteolin

wogonin

and baicalein may be potential candidate agents. Mitogen-activated protein kinase 14 (MAPK14)

recombinant V-Rel reticuloendotheliosis viral oncogene homolog A (RELA)

and tumor necrosis factor alpha (TNF-α) may be potent

ial therapeutic targets. Reactive oxygen species metabolic process and the apoptotic signaling pathway were the main biological processes. The p38MAPK/NF-κB signaling pathway might be the key signaling pathway activated by LGS against ARDS. Moreover

molecular docking demonstrated that luteolin

wogonin

and baicalein had a good binding affinity with MAPK14

RELA

and TNFα.

In vitro

experiments

LGS inhibited the expression and entry of p38 and p65 into the nucleation in human bronchial epithelial cells (HBE) cells induced by LPS

inhibited the inflammatory response and oxidative stress response

and inhibited HBE cell apoptosis (

P

<

0.05 or

P

<

0.01).

In vivo

experiments

LGS improved lung injury caused by ligation and puncture

reduced inflammatory responses

and inhibited the activation of p38MAPK and p65 (

P

<

0.05 or

P

<

0.01).

Conclusion:

2

LGS could reduce reactive oxygen species and inflammatory cytokine production by inhibiting p38MAPK/NF-κB signaling pathway

thus reducing apoptosis and attenuating ARDS.